The matrix metalloproteinases (MMPs) are a family of zinc containing endopeptidases which are capable of cleaving large biomolecules such as the collagens, proteoglycans and gelatins. Expression is upregulated by pro-inflammatory cytokines and/or growth factors. The MMP""s are secreted as inactive zymogens which, upon activation, are subject to control by endogenous inhibitors, for example, tissue inhibitor of metalloproteinases (TIMP) and xcex12-macroglobulin. Chapman, K. T. et al., J. Med. Chem. 36, 4293-4301 (1993); Beckett, R. P. et al., DDT 1, 16-26 (1996). The characterizing feature of diseases involving the enzymes appears to be a stoichiometric imbalance between active enzymes and endogenous inhibitors, leading to excessive tissue disruption, and often degradation. McCachren, S. S., Arthritis Rheum. 34, 1085-1093 (1993).
The discovery of different families of matrix metalloproteinase, their relationships, and their individual characteristics have been categorized in several reports. Emonard, H. et al., Cell Molec. Biol. 36, 131-153 (1990); Birkedal-Hansen, H., J. Oral Pathol. 17,445-451 (1988); Matrisian, L. M., Trends Genet. 6, 121-125 (1990); Murphy, G. J. P. et al., FEBS Lett. 289, 4-7 (1991); Matrisian, L. M., Bioessays 14, 455-463 (1992). Three groups of MMPs have been delineated: the collagenases which have triple helical interstitial collagen as a substrate, the gelatinases which are proteinases of denatured collagen and Type IV collagen, and the stromelysins which were originally characterized as proteoglycanases but have now been identified to have a broader characterized as proteoglycanases but have now been identified to have a broader proteolytic spectrum. Examples of specific collagenases include fibroblast collagenase (MMP-1), neutrophil collagenase (MMP-8), and collagenase 3 (MMP-13). Examples of gelatinases include 72 kDa gelatinase (gelatinase A; MMP-2) and 92 kDa gelatinase (gelatinase B; MMP-9). Examples of stromelysins include stromelysin 1 (MMP-3), stromelysin 2 (MMP-10) and matrilysin (MMP-7). Other MMPs which do not fit neatly into the above groups include metalloelastase (MMP-12), membrane-type MMP (MT-MMP or MMP-14) and stromelysin 3 (MMP-11). Beckett, R. P. et al., supra.
Over-expression and activation of MMPs have been linked with a wide range of diseases such as cancer; rheumatoid arthritis; osteoarthritis; chronic inflammatory disorders, such as emphysema and smoking-induced emphysema; cardiovascular disorders, such as atherosclerosis; corneal ulceration; dental diseases such as gingivitis and periodontal disease; and neurological disorders, such as multiple sclerosis. For example, in adenocarcinoma, invasive proximal gastric cells express the 72 kDa form of collagenase Type IV, whereas the noninvasive cells do not. Schwartz, G. K. et al., Cancer 73, 22-27 (1994). Rat embryo cells transformed by the Ha-ras and v-myc oncogenes or by Ha-ras alone are metastatic in nude mice and release the 92 kDa gelatinase/collagenase (MMP-9). Bernhard, E. J. et al., Proc. Natl. Acad. Sci. 91, 4293-4597 (1994). The plasma concentration of MMP-9 was significantly increased (P less than 0.01) in 122 patients with gastrointestinal tract cancer and breast cancer. Zucker, S. et al., Cancer Res. 53, 140-146 (1993). Moreover, intraperitoneal administration of batimastat, a synthetic MMP inhibitor, gave significant inhibition in the growth and metastatic spread and number of lung colonies which were produced by intravenous injection of the B16-BL6 murine melanoma in C57BL/6N mice. Chirivi, R. G. S. et al., Int. J. Cancer 58, 460-464 (1994). Over-expression of TIMP-2, the endogenous tissue inhibitor of MMP-2, markedly reduced melanoma growth in the skin of immunodeficient mice. Montgomery, A. M. P. et al., Cancer Res. 54, 5467-5473 (1994).
Accelerated breakdown of the extracellular matrix of articular cartilage is a key feature in the pathology of both rheumatoid arthritis and osteoarthritis. Current evidence suggests that the inappropriate synthesis of MMPs is the key event. Beeley, N. R. A. et al., Curr. Opin. Ther. Patents, 4(1), 7-16 (1994). The advent of reliable diagnostic tools have allowed a number of research groups to recognize that stromelysin is a key enzyme in both arthritis and joint trauma Beeley, N. R. A. et al., Id.; Hasty, K. A. et al., Arthr. Rheum. 33, 388-397 (1990). It has also been shown that stromelysin is important for the conversion of procollagenase to active collagenase. Murphy, G. et al., Biochem. J. 248, 265-268 (1987).
Furthermore, a range of MMPs can hydrolyse the membrane-bound precursor of the pro-inflammatory cytokine tumor necrosis factor xcex1 (TNF-xcex1). Gearing, A. J. H. et al., Nature 370, 555-557 (1994). This cleavage yields mature soluble TNF-xcex1 and the inhibitors of MMPs can block production of TNF-xcex1 both in vitro and in vivo. Gearing, A. J. H. et al., Id.; Mohler, K. M. et al., Nature 370, 218-220 (1994); McGeehan, G. M. et al., Nature 370, 558-561 (1994). This pharmacological action is a probable contributor to the antiarthritic action of this class of compounds seen in animal models. Beckett, R. P. et al., supra.
Stromelysin has been observed to degrade the xcex11-proteinase inhibitor which regulates the activity of enzymes such as elastase, excesses of which have been linked to chronic inflammatory disorders such as emphysema and chronic bronchitis. Beeley, N. R. A. et al., supra.; Wahl, R. C. et al., Annual Reports in Medicinal Chemistry 25, 177-184 (1990). In addition, a recent study indicates that MMP-12 is required for the development of smoking-induced emphysema in mice. Science, 277, 2002 (1997). Inhibition of the appropriate MMP may thus potentiate the inhibitory activity of endogenous inhibitors of this type.
High levels of mRNA corresponding to stromelysin have been observed in atherosclerotic plaques removed from heart transplant patients. Henney, A. M., et al., Proc. Natl. Acad. Sci. 88, 8154-8158 (1991). It is submitted that the role of stromelysin in such plaques is to encourage rupture of the connective tissue matrix which encloses the plaque. This rupture is in turn thought to be a key event in the cascade which leads to clot formation of the type seen in coronary thrombosis. MMP inhibition is thus a preventive measure for such thromboses.
Collagenase, stromelysin and gelatinase have been implicated in the destruction of the extracellular matrix of the cornea. This is thought to be an important mechanism of morbidity and visual loss in a number of ulcerative ocular diseases, particularly those following infection or chemical damage. Burns, F. R. et al., Invest. Opthalmol. and Visual Sci. 32, 1569-1575 (1989). The MMPs present in the eye during ulceration are derived either endogenously from infiltrating leucocytes or fibroblasts, or exogenously from microbes.
Collagenase and stromelysin activities have been identified in fibroblasts isolated from inflamed gingiva and the levels of enzyme have been correlated with the severity of the gingivitis observed. Beeley, N. R. A. et al., supra; Overall, C. M. et al., J. Periodontal Res. 22, 81-88 (1987).
Excessive levels of gelatinase-B in cerebrospinal fluid has been linked with incidence of multiple sclerosis and other neurological disorders. Beeley, N. R. A. et al., supra.; Miyazaki, K. et al., Nature 362, 839-841 (1993). The enzyme may play a key role in the demyelination of neurones and the breakdown of the blood brain barrier which occurs in such disorders.
The present invention provides a method of inhibiting matrix metallo-proteinases (MMPs) in a patient in need thereof comprising administering to the patient an effective matrix metalloproteinase inhibiting amount of the N-carboxymethyl substituted benzolactams of formula (1): 
wherein
A is selected from the group consisting of xe2x80x94OH and xe2x80x94NRRxe2x80x2;
wherein
R and Rxe2x80x2 are independently selected from the group consisting of hydrogen and C1-C6 alkyl or R and Rxe2x80x2 taken together with the nitrogen atom to which they are attached form a N-morpholino, N-piperidino, N-pyrrolidino, or N-isoindolyl;
R1 is selected from the group consisting of hydrogen, C1-C6 alkyl, xe2x80x94(CH2)axe2x80x94CO2R5, xe2x80x94(CH2)axe2x80x94C(O)NH2, xe2x80x94(CH2)4NH2, xe2x80x94(CH2)3xe2x80x94NHxe2x80x94C(NH)NH2, xe2x80x94(CH2)2xe2x80x94S(O)bxe2x80x94CH3, xe2x80x94CH2xe2x80x94OH,
xe2x80x94CH(OH)CH3, xe2x80x94CH2xe2x80x94SH, xe2x80x94(CH2)dxe2x80x94Ar1, and xe2x80x94CH2xe2x80x94Ar2;
wherein
a is 1 or 2;
b is 0, 1, or 2;
d is an integer from 0 to 4;
R5 is selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
Ar1 is a radical selected from the group consisting of 
wherein
R6 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, hydroxy, and C1-C4 alkoxy;
R7 is selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
Ar2 is a radical selected from the group consisting of 
R2 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, hydroxy C1-C4 alkyl, and C1-C4 alkoxy;
R3 is selected from the group consisting of C1-C6 alkyl, xe2x80x94(CH2)mxe2x80x94W, xe2x80x94(CH2)pxe2x80x94Ar3, xe2x80x94(CH2)kxe2x80x94CO2R9, xe2x80x94(CH2)mxe2x80x94NR8xe2x80x2SO2xe2x80x94Y1, and xe2x80x94(CH2)m-Z-Q
wherein
m is an integer from 2 to 8;
p is an integer from 0-10;
k is an integer from 1 to 9;
W is phthalimido;
Ar3 is selected from the group consisting of 
wherein
R23 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
R8xe2x80x2 is hydrogen or C1-C6 alkyl;
R9 is hydrogen or C1-C6 alkyl;
Y1 is selected from the group consisting of hydrogen, xe2x80x94(CH2)jxe2x80x94Ar4, and xe2x80x94N(R24)2 
wherein
j is 0 or 1;
R24 each time selected is independently hydrogen or C1-C6 alkyl or are taken together with the nitrogen to which they are attached to form N-morpholino, N-piperidino, N-pyrrolidino, or N-isoindolyl;
Ar4 is 
wherein
R25 is from 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
Z is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94NR8xe2x80x94, xe2x80x94C(O)NR8xe2x80x94, xe2x80x94NR8C(O)xe2x80x94, xe2x80x94NR8C(O)NHxe2x80x94, xe2x80x94NR8C(O)Oxe2x80x94, and xe2x80x94OC(O)NHxe2x80x94;
wherein
R8 is hydrogen or C1-C6 alkyl;
Q is selected from the group consisting of hydrogen, xe2x80x94(CH2)nxe2x80x94Y2, and xe2x80x94(CH2)xY3;
wherein
n is an integer from 0 to 4;
Y2 is selected from the group consisting of hydrogen, xe2x80x94(CH2)hxe2x80x94Ar5 and xe2x80x94(CH2)txe2x80x94C(O)OR27 
wherein
Ar5 is selected from the group consisting of 
wherein
R26 is from 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
h is an integer from 0 to 6;
t is an integer from 1 to 6;
R27 is hydrogen or C1-C6 alkyl;
x is an integer from 2 to 4;
Y3 is selected from the group consisting of xe2x80x94N(R28)2, N-morpholino, N-piperidino, N-pyrrolidino, and N-isoindolyl;
wherein
R28 each time taken is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R4 is selected from the group consisting of hydrogen, xe2x80x94C(O)R10, xe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94K and xe2x80x94S-G
wherein
R10 is selected from the group consisting of hydrogen, C1-C4 alkyl, phenyl, and benzyl;
q is 0, 1, or 2;
K is selected from the group consisting of 
wherein
V is selected from the group consisting of a bond, xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94S(O)rxe2x80x94, xe2x80x94NR21xe2x80x94, and xe2x80x94NC(O)R22;
wherein
r is 0, 1, or 2;
R21 is selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
R22 is selected from the group consisting of hydrogen, xe2x80x94CF3, C1-C10 alkyl, phenyl, and benzyl;
R11 each time taken is independently selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
G is selected from the group consisting of 
wherein
w is an integer from 1 to 3;
R12 is selected from the group consisting of hydrogen, C1-C6 alkyl, xe2x80x94CH2CH2S(O)uCH3, and benzyl;
wherein
u is 0, 1, or 2;
R13 is selected from the group consisting of hydrogen, hydroxy, amino, C1-C6 alkyl, N-methylamino, N,N-dimethylamino, xe2x80x94CO2R17, and xe2x80x94OC(O)R18;
wherein
R17 is hydrogen, xe2x80x94CH2Oxe2x80x94C(O)C(CH3)3, C1-C4 alkyl, benzyl, or diphenylmethyl;
R18 is hydrogen, C1-C6 alkyl or phenyl;
R14 is 1 or 2 substituents independently selected from the group consisting of hydrogen, C1-C4 alkyl, C1-C4 alkoxy, or halogen;
V1 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94NHxe2x80x94;
V2 is selected from the group consisting of xe2x80x94Nxe2x80x94 and xe2x80x94CHxe2x80x94,
V3 is selected from the group consisting of a bond and xe2x80x94C(O)xe2x80x94;
V4 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94NR19xe2x80x94, and xe2x80x94NC(O)R20xe2x80x94;
wherein
R19 is hydrogen, C1-C4 alkyl, or benzyl;
R20 is hydrogen, xe2x80x94CF3, C1-C10 alkyl, or benzyl;
R15 is selected from the group consisting of hydrogen, C1-C6 alkyl and benzyl;
R16 is selected from the group consisting of hydrogen and C1-C4 alkyl; and stereoisomers, pharmaceutically acceptable salt, and hydrate thereof.
Novel N-carboxymethyl substituted benzolactams are encompassed by formula (1). Some of these novel compounds are described by of formula (1a), below, which is encompassed by formula (1). The present invention provides novel N-carboxymethyl substituted benzolactams of formula (1a): 
wherein
Aa is xe2x80x94NRRxe2x80x2;
wherein
R and Rxe2x80x2 are independently selected from the group consisting of hydrogen and C1-C6 alkyl or R and Rxe2x80x2 taken together with the nitrogen atom to which they are attached form a N-morpholino, N-piperidino, N-pyrrolidino, or N-isoindolyl;
R1 is selected from the group consisting of hydrogen, C1-C6 alkyl, (CH2)axe2x80x94CO2R5, xe2x80x94(CH2)axe2x80x94C(O)NH2, xe2x80x94(CH2)4NH2, xe2x80x94(CH2)3xe2x80x94NHxe2x80x94C(NH)2, xe2x80x94(CH2)2xe2x80x94S(O)bxe2x80x94CH3, xe2x80x94CH2xe2x80x94OH,
xe2x80x94CH(OH)CH3, xe2x80x94CH2xe2x80x94SH, xe2x80x94(CH2)dxe2x80x94Ar1, and xe2x80x94CH2xe2x80x94Ar2;
wherein
a is 1 or 2;
b is 0, 1, or 2;
d is an integer from 0 to 4;
R5 is selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
Ar1 is a radical selected from the group consisting of 
wherein
R6 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, hydroxy, and C1-C4 alkoxy;
R7 is selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
Ar2 is a radical selected from the group consisting of 
R2 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, hydroxy C1-C4 alkyl, and C1-C4 alkoxy;
R3 is selected from the group consisting of C1-C6 alkyl, xe2x80x94(CH2)mxe2x80x94W, xe2x80x94(CH2)pxe2x80x94Ar3, xe2x80x94(CH2)kxe2x80x94CO2R9, xe2x80x94(CH2)mxe2x80x94NR8xe2x80x2SO2xe2x80x94Y1, and xe2x80x94(CH2)m-Z-Q
wherein
m is an integer from 2 to 8;
p is an integer from 0-10;
k is an integer from 1 to 9;
W is phthalimido;
Ar3 is selected from the group consisting of 
wherein
R23 is from 1 to 2 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
R8xe2x80x2 is hydrogen or C1-C6 alkyl;
R9 is hydrogen or C1-C6 alkyl;
Y1 is selected from the group consisting of hydrogen, xe2x80x94(CH2)jxe2x80x94Ar4, and xe2x80x94N(R24)2 
wherein
j is 0 or 1;
R24 each time selected is independently hydrogen or C1-C6 alkyl or are taken together with the nitrogen to which they are attached to form N-morpholino, N-piperidino, N-pyrrolidino, or N-isoindolyl;
Ar4 is 
wherein
R25 is from 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
Z is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94NR8xe2x80x94, xe2x80x94C(O)NR8xe2x80x94, xe2x80x94NR8C(O)xe2x80x94, xe2x80x94NR8C(O)NHxe2x80x94, xe2x80x94NR8C(O)Oxe2x80x94, and xe2x80x94OC(O)NHxe2x80x94;
wherein
R8 is hydrogen or C1-C6 alkyl;
Q is selected from the group consisting of hydrogen, xe2x80x94(CH2)nxe2x80x94Y2, and xe2x80x94(CH2)xY3;
wherein
n is an integer from 0 to 4;
Y2 is selected from the group consisting of hydrogen, xe2x80x94(CH2)hxe2x80x94Ar5 and xe2x80x94(CH2)txe2x80x94C(O)OR27 
wherein
Ar5 is selected from the group consisting of 
wherein
R26 is from 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, C1-C4 alkyl, and C1-C4 alkoxy;
h is an integer from 0 to 6;
t is an integer from 1 to 6;
R27 is hydrogen or C1-C6 alkyl;
x is an integer from 2 to 4;
Y3 is selected from the group consisting of xe2x80x94N(R28)2, N-morpholino, N-piperidino, N-pyrrolidino, and N-isoindolyl;
wherein
R28 each time taken is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R4 is selected from the group consisting of hydrogen, xe2x80x94C(O)R10, xe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94K and xe2x80x94S-G
wherein
R10 is selected from the group consisting of hydrogen, C1-C4 alkyl, phenyl, and benzyl;
q is 0, 1, or 2;
K is selected from the group consisting of 
wherein
V is selected from the group consisting of a bond, xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94S(O)rxe2x80x94, xe2x80x94NR21xe2x80x94, and xe2x80x94NC(O)R22;
wherein
r is 0, 1, or 2;
R21 is selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
R22 is selected from the group consisting of hydrogen, xe2x80x94CF3, C1-C10 alkyl, phenyl, and benzyl;
R11 each time taken is independently selected from the group consisting of hydrogen, C1-C4 alkyl, and benzyl;
G is selected from the group consisting of 
wherein
w is an integer from 1 to 3;
R12 is selected from the group consisting of hydrogen, C1-C6 alkyl, xe2x80x94CH2CH2S(O)uCH3, and benzyl;
wherein
u is 0, 1, or 2;
R13 is selected from the group consisting of hydrogen, hydroxy, amino, C1-C6 alkyl, N-methylamino, N,N-dimethylamino, xe2x80x94CO2R17, and xe2x80x94OC(O)R18;
wherein
R17 is hydrogen, xe2x80x94CH2Oxe2x80x94C(O)C(CH3)3, C1-C4 alkyl, benzyl, or diphenylmethyl;
R18 is hydrogen, C1-C6 alkyl or phenyl;
R14 is 1 or 2 substituents independently selected from the group consisting of hydrogen, C1-C4 alkyl, C1-C4 alkoxy, or halogen;
V1 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94NHxe2x80x94;
V2 is selected from the group consisting of xe2x80x94Nxe2x80x94 and xe2x80x94CHxe2x80x94;
V3 is selected from the group consisting of a bond and xe2x80x94C(O)xe2x80x94;
V4 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94NR19xe2x80x94, and xe2x80x94NC(O)R20xe2x80x94;
wherein
R19 is hydrogen, C1-C4 alkyl, or benzyl;
R20 is hydrogen, xe2x80x94CF3, C1-C10 alkyl, or benzyl;
R15 is selected from the group consisting of hydrogen, C1-C6 alkyl and benzyl;
R16 is selected from the group consisting of hydrogen and C1-C4 alkyl; and stereoisomers, pharmaceutically acceptable salt, and hydrate thereof.
In addition, the present invention provides a composition comprising an assayable amount of a compound of formula (1a) in admixture or otherwise in association with an inert carrier. The present invention also provides a pharmaceutical composition comprising an effective matrix metallo-proteinases inhibitory amount of a compound of formula (1a) in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.
As is appreciated by one of ordinary skill in the art the compounds of formula (1) exist as stereoisomers. Specifically, it is recognized that they exist as stereoisomers at the point of attachment of the substituents R1, R3, and xe2x80x94SR4, R12, and xe2x80x94NHR15 and at the point of attachment of the group xe2x80x94NHC(O)CH(R3)(SR4) to the benzolactam. Where indicated the compounds, whether of formula (1), starting materials, or intermediates, follow either the (+)- and (xe2x88x92)-designation for optical rotation, the (D)- and (L)-designation of relative stereochemistry, or the Cahn-Ingold-Prelog designation of (R)- and (S)- for the stereochemistry of at specific positions in the compounds represented by formula (1) and intermediates thereof. Any reference in this application to one of the compounds of the formula (1) is meant to encompass either specific stereoisomers or a mixture of stereoisomers.
The specific stereoisomers can be prepared by stereospecific synthesis using enantiomerically pure or enantiomerically enriched starting materials which are well known in the art. The specific stereoisomers of amino acid starting materials are commercially available or can be prepared by stereospecific synthesis as is well known in the art or analogously known in the art, such as D. A. Evans, et al. J. Am. Chem. Soc., 112, 4011-4030 (1990); S. Ikegami et al. Tetrahedron, 44, 5333-5342 (1988); W. Oppolzer et al. Tet. Lets. 30, 6009-6010 (1989); Synthesis of Optically Active xcex1-Amino-Acids, R. M. Williams (Pergamon Press, Oxford 1989); M. J. O""Donnell ed.: xcex1-Amino-Acid Synthesis, Tetrahedron Symposia in print, No. 33, Tetrahedron 44, No. 17 (1988); U. Schxc3x6llkopf, Pure Appl. Chem. 55, 1799 (1983); U. Hengartner et al. J. Org. Chem., 44, 3748-3752 (1979); M. J. O""Donnell et al. Tet. Lets., 2641-2644 (1978); M. J. O""Donnell et al. Tet. Lets. 23, 4255-4258 (1982); M. J. O""Donnell et al. J. Am. Chem. Soc., 110, 8520-8525(1988).
The specific stereoisomers of either starting materials or products can be resolved and recovered by techniques known in the art, such as chromatography on chiral stationary phases, enzymatic resolution, or fractional recrystallization of addition salts formed by reagents used for that purpose. Useful methods of resolving and recovering specific stereoisomers are known in the art and described in Stereochemistry of Organic Compounds, E. L. Eliel and S. H. Wilen, Wiley (1994) and Enantiomers, Racemates, and Resolutions, J. Jacques, A. Collet, and S. H. Wilen, Wiley (1981).
As used in this application:
a) the term xe2x80x9chalogenxe2x80x9d refers to a fluorine atom, chlorine atom, bromine atom, or iodine atom;
b) the term xe2x80x9cC1-C6 alkylxe2x80x9d refers to a branched or straight chained alkyl radical containing from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, etc.;
c) the term xe2x80x9cC1-C4 alkylxe2x80x9d refers to a saturated straight or branched chain alkyl group containing from 1-4 carbon atoms and includes methyl, ethyl, propyl, isopropyl, n-butyl, s-butyl, isobutyl, and t-butyl;
d) the term xe2x80x9cC1-C10 alkylxe2x80x9d refers to a branched or straight chained alkyl radical containing from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.;
e) the term xe2x80x9cC1-C4 alkoxyxe2x80x9d refers to a straight or branched alkoxy group containing from 1 to 4 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy, etc.;
f) the designation xe2x80x9cxe2x80x9d refers to a bond for which the stereochemistry is not designated;
g) the designation xe2x80x9cxe2x80x9d refers to a bond that protrudes forward out of the plane of the page.
h) the designation xe2x80x9cxe2x80x9d refers to a bond that protrudes backward out of the plane of the page.
i) as used in the examples and preparations, the following terms have the meanings indicated: xe2x80x9cgxe2x80x9d refers to grams, xe2x80x9cmgxe2x80x9d refers to milligrams, xe2x80x9cxcexcgxe2x80x9d refers to micrograms, xe2x80x9cmolxe2x80x9d refers to moles, xe2x80x9cmmolxe2x80x9d refers to millimoles, xe2x80x9cnmolexe2x80x9d refers to nanomoles, xe2x80x9cLxe2x80x9d refers to liters, xe2x80x9cmLxe2x80x9d or xe2x80x9cmlxe2x80x9d refers to milliliters, xe2x80x9cxcexcLxe2x80x9d refers to microliters, xe2x80x9cxc2x0 C.xe2x80x9d refers to degrees Celsius, xe2x80x9cRfxe2x80x9d refers to retention factor, xe2x80x9cmpxe2x80x9d refers to melting point, xe2x80x9cdecxe2x80x9d refers to decomposition, xe2x80x9cbpxe2x80x9d refers to boiling point, xe2x80x9cmm of Hgxe2x80x9d refers to pressure in millimeters of mercury, xe2x80x9ccmxe2x80x9d refers to centimeters, xe2x80x9cmmxe2x80x9d refers to nanometers, xe2x80x9cbrinexe2x80x9d refers to a saturated aqueous sodium chloride solution, xe2x80x9cMxe2x80x9d refers to molar, xe2x80x9cmMxe2x80x9d refers to millimolar, xe2x80x9cxcexcMxe2x80x9d refers to micromolar, xe2x80x9cnMxe2x80x9d refers to nanomolar, xe2x80x9cHPLCxe2x80x9d refers to high performance liquid chromatography, xe2x80x9cHRMSxe2x80x9d refers to high resolution mass spectrum, xe2x80x9cDMFxe2x80x9d refers to dimethylformamide, xe2x80x9cxcexcCixe2x80x9d refers to microcuries, xe2x80x9ci.p.xe2x80x9d refers to intraperitoneally, xe2x80x9ci.v.xe2x80x9d refers to intravenously, and xe2x80x9cDPMxe2x80x9d refers to disintegrations per minute;
j) for substituent Z, the designations xe2x80x94C(O)NR8xe2x80x94, xe2x80x94NR8C(O)xe2x80x94, xe2x80x94NR8C(O)NHxe2x80x94, xe2x80x94NR8C(O)Oxe2x80x94, and xe2x80x94OC(O)NHxe2x80x94 refer to the functionalities represented, respectively, by the following formulae showing the attachment of the group (Q): 
these designations are referred to hereinafter as amido, amide, urea, N-carbamoyl, and O-carbamoyl, respectively;
k) the term xe2x80x9cpharmaceutically acceptable salts thereof refers to either an acid addition salt or a basic addition salt.
The expression xe2x80x9cpharmaceutically acceptable acid addition saltsxe2x80x9d is intended to apply to any non-toxic organic or inorganic acid addition salt of the base compounds represented by formula (1) or any of its intermediates. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulphuric, and phosphoric acid and acid metal salts such as sodium monohydrogen orthophosphate, and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include the mono-, di-, and tricarboxylic acids. Illustrative of such acids are for example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, salicyclic, 2-phenoxybenzoic, p-toluenesulfonic acid, and sulfonic acids such as methane sulfonic acid and 2-hydroxyethane sulfonic acid. Such salts can exist in either a hydrated or substantially anhydrous form. In general, the acid addition salts of these compounds are soluble in water and various hydrophilic organic solvents, and which in comparison to their free base forms, generally demonstrate higher melting points.
The expression xe2x80x9cpharmaceutically acceptable basic addition saltsxe2x80x9d is intended to apply to any non-toxic organic or inorganic basic addition salts of the compounds represented by formula (1) or any of its intermediates. Illustrative bases which form suitable salts include alkali metal or alkaline-earth metal hydroxides such as sodium, potassium, calcium, magnesium, or barium hydroxides; ammonia, and aliphatic, alicyclic, or aromatic organic amines such as methylamine, dimethylamine, trimethylamine, and picoline.
As with any group of structurally related compounds which possess a particular utility, certain groups and configurations of substituents are preferred for the compounds of formula (1). Preferred embodiments are given below:
The compounds in which R1 is selected from the group consisting of C1-C6 alkyl and xe2x80x94(CH2)dAr1 are preferred;
The compounds in which R1 is xe2x80x94(CH2)dAr1 and Ar1 is phenyl or substituted phenyl are more preferred;
Compounds in which R4 is selected from the group consisting of hydrogen, xe2x80x94C(O)R10 and xe2x80x94S-G are preferred;
Compounds in which R4 is hydrogen, are more preferred;
Compounds in which R4 is xe2x80x94C(O)R10 and R10 is C1-C4 alkyl more preferred;
Compounds in which A is xe2x80x94OH are preferred; and
Compounds in which A is xe2x80x94NRRxe2x80x2 wherein R is hydrogen and Rxe2x80x2 is methyl are preferred.
Examples of compounds encompassed by formula (1) and (1a) of the present invention include the following. It is understood that the examples encompass all of the isomers of the compound and mixtures thereof. This list is meant to be representative only and is not intended to limit the scope of the invention in any way: 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, (S)xe2x80x94N-1-(2-methylpropyl)-2-(thio)-ethylamine, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, (R)-1-(2-methylpropyl)-2-(thio)-ethylamine, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4 methyl-valeric acid, L-cysteine ethyl ester, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, N-acetyl-L-cysteine ethyl ester, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, L-cysteine, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, benzylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid ethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, disulfide, 2-hydroxyethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, t-butyl ester, 2-pyridylmethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, 2-thioacetic acid morpholine carboxamide, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, t-butyl ester, benzylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid t-butyl ester, ethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, t-butyl ester, disulfide, 2-hydroxyethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, t-butyl ester, 2-pyridylmethylthio, disulfide; 2-(4-(2-Thio-3-phenyl-propionyl-amino)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, t-butyl ester, 2-thioacetic acid morpholine carboxamide, disulfide.
As used herein the term xe2x80x9camino acidxe2x80x9d refers to naturally occurring amino acids as well as non-naturally occurring amino acids having substituents encompassed by R1 and R2 as described above. The naturally occurring amino acids included are glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, histidine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, ornithine, and lysine. Non-naturally occurring amino acids within the term xe2x80x9camino acid,xe2x80x9d include without limitation, the D-isomers of the naturally occurring amino acids, norleucine, norvaline, alloisoleucine, t-butylglycine, methionine sulfoxide, and methionine sulfone. Other non-naturally occurring amino acids within the term xe2x80x9camino acid,xe2x80x9d include without limitation phenylalanines, phenylglycines, homophenylalanines, 3-phenylpropylglycines, 4-phenylbutylglycines; each including those substituted by R6 and R6 as described above; and 1-naphthylalanines and 2-naphthylalanines; including those substituted by R7 and R7xe2x80x2 as described above.
The compounds of formula (1) can be prepared by utilizing techniques and procedures well known and appreciated by one of ordinary skill in the art. To illustrate, general synthetic schemes for preparing starting material, intermediates, and the compounds of formula (1) are set forth below. In the reaction schemes below, the reagents and starting materials are readily available to one of ordinary skill in the art and all substituents are as previously defined unless otherwise-indicated. 
In Reaction Scheme A, step 1, a compound of formula (3) is coupled with a with an appropriate acid derivative of formula (2) to give a compound of formula (4). Such coupling reactions are well known in the art.
An appropriate compound of structure (3) in one in which R2 is as desired in the final compound of formula (1), R1 as desired in the final product of formula (1) or gives rise after deprotection to R1 as desired in the final product of formula (1), and Axe2x80x2 is A or gives rise to A upon deprotection and modification, if desired, to A as desired in the final compound of formula (1). Appropriate compounds of formula (3) can be prepared by methods described herein and by methods well known and appreciated in the art as described in PCT International Publication Number WO 95/21854, published Aug. 17, 1995 and European Patent Application Publication Number 0 599 444 A1, published Jun. 1, 1994.
An appropriate compound of formula (2) is one in which R3xe2x80x2 is R3 as desired in the final product of formula (1) or gives rise after deprotection to R3 as desired in the final product of formula (1) and Y is a protected thio substituent or Y may be a protected hydroxy substituent or bromo which gives rise upon selective deprotection and displacement or displacement and further deprotection and/or elaboration, if required, to xe2x80x94SR4 as desired in the final product of formula (1). Alternately, an appropriate compound of formula (2) may also be one in which R3xe2x80x2 gives rise to R3xe2x80x3 which, upon farther reaction, gives rise R3 as desired in the final product of formula (1), as described in Reaction Scheme B, and Y is a protected thio substituent. In addition, an appropriate compound of formula (2) may also be one in which the stereochemistry at the R3xe2x80x2 and Y bearing carbon is as desired in the final product of formula (1) or gives rise after displacement to the stereochemistry as desired at that carbon in the final product of formula (1).
The use and selection of appropriate protecting groups is within the ability of those skilled in the art and will depend upon compound of formula (2) to be protected, the presence of other protected amino acid residues, other protecting groups, and the nature of the particular R3 and/or R4 group(s) ultimately being introduced. Compounds of formula (2) in which Y is bromo and protected thio are commercially available or can be prepared utilizing materials, techniques, and procedures well known and appreciated by one of ordinary skill in the art or described herein. See PCT Application WO 96/11209, published Apr. 8, 1996. Examples commercially available compounds of formula (2) in which Y is bromo include 2-bromopropionic acid, 2-bromobutyric acid, 2-bromovaleric acid, 2-bromohexanoic acid, 6-(benzoylamino)-2-bromohexanoic acid, 2-bromoheptanoic acid, 2-bromooctanoic acid, 2-bromo-3-methylbutyric acid, 2-bromoisocaproic acid, 2-bromo-3-(5-imidazoyl)proionic acid, (R)-(+)-2-bromopropionic acid, (S)-(xe2x88x92)-2-bromopropionic acid.
For example, a compound of formula (3) is contacted in a coupling reaction with a compound of formula (2). The compound of formula (2) may be converted to an activated intermediate; such as and acid chloride, an anhydride; a mixed anhydride of aliphatic carboxylic acid, such as formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, 2-ethylbutyric acid, trichloroacetic acid, trifluoroacetic acid, and the like; of aromatic carboxylic acids, such as benzoic acid and the like; of an activated ester, such as phenol ester, p-nitrophenol ester, 2,4-dinitrophenol ester, pentafluorophenol ester, pentachlorophenol ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 1-hydroxy-1H-benztriazole ester, O-azabenztriazoyl-N,N,Nxe2x80x2,Nxe2x80x2-tetramethyluronium hexafluoro-phosphate and the like; activated amide, such as imidazole, dimethylpyrazole, triazole, or tetrazole; or an intermediate formed in the presence of coupling agents, such as dicyclohexylcarbodiimide or 1-(3-dimethyaminopropyl)-3-ethylcarbodiimide. The reaction is carried out in a suitable solvent, such as dichloromethane, chloroform, tetrahydrofuran, or dimethylformafide. The reaction generally is carried out at temperatures of from about xe2x88x9220xc2x0 C. to the refluxing temperature of the solvent and generally requires 1 to 48 hours. The product can be isolated and purified by techniques well known in the art, such as filtration, extraction, evaporation, chromatography, and recrystallization.
In Reaction Scheme A, step 2, a compound of formula (4) in which Y is hydroxy or bromo gives rise to a compound of formula (5) in which Y is protected thio.
A compound of formula (4) in which Y is hydroxy (obtained from protected hydroxy compounds of formula (2)) undergoes a displacement reaction with an appropriate thio introducing reagent by the method of Mitsunobu to give a compound of formula (5) in which Y is a protected thio substituent. For example, a compound of formula (4) in which Y is hydroxy reacts with thioacetic acid or thiobenzoic acid, triphenylphosphine, and diethylazodicarboxylate in a suitable aprotic solvent, such as tetrahydrofuran to give a compound of formula (5) in which Y is thioacetyl or thiobenzoyl. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, lyophilization, chromatography, and recrystallization.
A compound of formula (4) in which Y is bromo undergo a displacement reaction with an appropriate thio introducing reagent to give a compound of formula (5) in which Y is protected thio substituent which gives rise upon deprotection and subsequent elaboration, if desired, the xe2x80x94SR4 as desired in the final compound of formula (1).
For example, a solution of p-methoxybenzylmercaptan in a suitable organic solvent such as dimethylformamide is degassed and treated with a suitable base such as sodium hydride. After about 1 to 2 hours, a solution of a compound of formula (4) in which Y is bromo is added. The reaction may benefit from the addition of a suitable catalyst, such as tetra-n-butylammonium iodide. The reaction mixture is carried out for 1 to 25 hours at temperatures ranging from 0xc2x0 C. to about 100xc2x0 C. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, lyophilization, chromatography, and recrystallization.
In Reaction Scheme A, step 3, a compound of formula (5) in which Y is protected thio undergoes selective deprotection to give a compound of formula (6). Protected thio substituents include thioesters, such as thioacetyl or thiobenzoyl, thioethers, such as thiobenzyl, thio-4-methoxybenzyl, thiotriphenylmethyl, or thio-t-butyl, or unsymmetrical sulfides, such as dithioethyl or dithio-t-butyl. The use and selective removal of thio protecting groups is well known and appreciated in the art and described in Protective Groups in Organic Synthesis, Theodora W. Greene (Wiley-Interscience, 2nd Edition, 1991).
Alternately, in Reaction Scheme A, step 4, a compound of formula (4) in which Y is protected thio is selectively deprotected to give a compound of formula (6), as described above, in Reaction Scheme A, step 3.
In Reaction Scheme A, step 5, a compound of formula (6) undergoes a undergoes modification reaction to give a compound of formula (7). Such modification reactions include, thiol esterification and disulfide formation.
Compounds of formula (7) in which R4 is xe2x80x94C(O)R10 or xe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94X group can be synthesized by thiol esterifications according to techniques well known and appreciated by one of ordinary skill in the art, such as those disclosed in U.S. Pat. No. 5,424,425, issued Jun. 13, 1995.
For example, in a thiol esterification a compound of formula (6) is contacted with about an equimolar amount of an appropriate acid, such as HOxe2x80x94C(O)R10 or HOxe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94X in the presence of a suitable coupling agent to give a compound of formula (6) in which R4 is xe2x80x94C(O)R10 or xe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94X. The reaction is carried out in the presence of a coupling agent such as 2-fluoro-1-methylpyridinium p-toluenesulfate, (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), carbonyldiimidazole, (1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, or diethylcyanophosphonate in a suitable aprotic solvent such as methylene chloride. The reaction is generally carried out at temperature of between xe2x88x9220xc2x0 C. and the boiling point of the solvent. Generally, the reaction requires 1 to 24 hours. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, trituration, lyophilization, chromatography, and recrystallization.
Compounds of formula (7) in which R4 is xe2x80x94S-G group can be synthesized according to techniques well known and appreciated by one of ordinary skill in the art, as disclosed in PCT Application No. WO 95/21839, published Aug. 17, 1995 and U.S. Pat. No. 5,491,143, issued Feb. 13, 1996, and U.S. Pat. No. 5,731,306, issued Mar. 24, 1998, and Roques, B. P. et al., J. Med. Chem. 33, 2473-2481 (1992).
For example, in a disulfide formation a compound of formula (6) is contacted with an appropriate compound of formula (8). 
An appropriate compound of formula (8) is one which gives G as desired in the final product of formula (1) or gives rise upon deprotection to G as is desired in the final product of formula (1). In addition, the compound of formula (8) may have stereochemistry as desired in the final product of formula (1). The reaction is carried out in a suitable solvent, such as ethanol, methanol, dichloromethane, or mixtures of ethanol or methanol and dichlorornethane. The solvent is degassed by passing a stream of nitrogen gas through it for 15 minutes before the reaction is carried out. The reaction is carried out using from 1.0 to 4.0 molar equivalents of an appropriate compound of formula (8). The reaction is carried out at temperatures of from 0xc2x0 C. to the refluxing temperature of the solvent, with a temperature of 10xc2x0 C. to 30xc2x0 C. being preferred. The reaction generally requires from 1 to 48 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
In Reaction Scheme A, step 6, a compound of formula (4) in which Y is hydroxy or bromo is displaced by an appropriate thiol, HSR4, by the methods described in Reaction. Scheme A, step 2, to give a compound of formula (7). In Reaction Scheme A, step 6, an appropriate thiol HSR4 is one which gives R4 as desired in the final product of formula (1) or gives rise to R4 as desired in the final product of formula (1). Also in Reaction Scheme A, step 6, a compound of formula (4) in which Y is bromo can be displaced by an appropriate thio ester, Ph3Sxe2x80x94C(O)xe2x80x94(CH2)qxe2x80x94X by techniques well known and appreciated in the art, as disclosed in U.S. Pat. No. 5,424,425, issued Jun. 13, 1995.
In Reaction Scheme A, a compound of formula (5), (6), or (7) is optionally deprotected to give a compound of formula (1). Such deprotection reactions are well known appreciated in the art and may include selective deprotections of protecting groups on Axe2x80x2, R1, R2, R3, and R4, as required to give the desired compound of formula (1). 
In Reaction Scheme B, step 1, an appropriate compound of formula (5) is deprotected, hydrolyzed, or reduced to give a compound of formula (9). In Reaction Scheme B, step 1, an appropriate compound of formula (5) is one in which Axe2x80x2 is A or gives rise to A upon deprotection and modification, if desired, in the final product of formula (1) and R1, and R2 are as desired in the final product of formula (1) or give rise upon deprotection to R1 and/or R2 as desired in the final product of formula (1). In Reaction Scheme B, step 1, an appropriate compound of formula (5) is one in which R3xe2x80x2 gives rise to a compound of formula (9) in which R3xe2x80x3 is R3 as desired in the final product of formula (1) or undergoes flirter derivitization (step 2) to give a compound of formula (10) in which R3 is as desired in the final product of formula (1). In Reaction Scheme B, step 1, an appropriate compound of formula (5) is one in which Y is xe2x80x94SR4 as desired in the final compound of formula (1) or Y gives rise upon deprotection (step 3) and further functionalization (step 4) or deprotection (step 5) to give xe2x80x94SR4 as desired in the final product of formula (1). In Reaction Scheme B, the use of compounds of formula (5) in which Y is a protected thio group, such as thioacetyl, thiobenzoyl, 4-methoxybenzylthio or t-butylthio is preferred.
For example, in a deprotection a compound of formula (5) in which R3xe2x80x2 is xe2x80x94(CH2)mxe2x80x94W (phthalimido group) is contacted with a molar excess of hydrazine monohydrate to give a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 in which R8 is hydrogen. The reaction is typically carried out in a protic organic solvent, such as methanol or ethanol. The reaction is generally carried out at room temperature for a period of time ranging from 5-24 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
Alternately, for example, in a deprotection of a compound of formula (5) in which R3xe2x80x2 is xe2x80x94(CH2)mxe2x80x94NR8-t-Boc is contacted with a molar excess of a suitable acid to give a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8. The reaction is typically carried out in a organic solvent, such as methanol, ethanol, ethyl acetate, diethyl ether, or dioxane. Suitable acids for this reaction are well known in the art, including hydrochloric acid, hydrobromic acid, trifluoroacetic acid, and methanesulfonic acid. The Keaction is generally carried out at temperatures from about 0xc2x0 C. to about room temperature for a period of time ranging from 1-10 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
For example, in a hydrolysis a compound of formula (5) in which R3xe2x80x2 is xe2x80x94(CH2)mxe2x80x94C(O)OPg3 and Pg3 is methyl or ethyl is contacted with about 1 to 2 molar equivalents of lithium hydroxide, sodium hydroxide, or potassium hydroxide to give a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94CO2H. The reaction is carried out in a suitable solvent, such as methanol, ethanol methanol/water mixtures, ethanol/water mixtures, or tetrahydrofuran/water mixtures and generally requires 1 to 24 hours. The reaction is carried out at temperatures of from about 0xc2x0 C. to the refluxing temperature of the solvent. The resulting acid is isolated and purified by techniques well known in the art, such as acidification, extraction, evaporation, and precipitation and can be purified by trituration, precipitation, chromatography, and recrystallization.
For example, in a reduction a compound of formula (5) in which R3xe2x80x2 is xe2x80x94(CH2)mxe2x88x921xe2x80x94CO2Pg3 in which Pg3 is methyl or ethyl is contacted with a suitable reducing agent, such as lithium borohydride, lithium aluminum hydride, diisobutylaluminum hydride, 9-borabicyclo(3.3.1)nonane, preferably lithium borohydride to provide a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x88x921xe2x80x94CH2OH. The reaction is carried out in a suitable solvent, such as dichloromethane, tetrahydrofuran, or toluene, with tetrahydrofuran being preferred. The reaction is carried out at a temperature of from about xe2x88x9230xc2x0 C. to about 50xc2x0 C. and generally requires from 2 to 12 hours. The product can be isolated by quenching, extraction, evaporation, and precipitation and can be purified by trituration, chromatography, and recrystallization.
In Reaction Scheme B, step 2, a compound of formula (9) undergoes a derivitization reaction to give a compound of formula (10) in which R3 is as desired in the final product of formula (1). Such derivitization reactions include hydrolysis of esters and ester formations as are well known in the art, ether formation, amine alkylation, formation of amides, urea formation, carbamate formation, and formation of sulfonamide.
For example, in an ether formation a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x88x921xe2x80x94CH2OH is contacted with 1 to 10 molar equivalents of a suitable akylating agent to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is xe2x80x94Oxe2x80x94. A suitable alkylating agent is one which transfers Q or protected Q as desired in the final product of formula (1), such as benzyl bromide, benzyl chloride, substituted benzyl bromide, substituted benzyl chloride, ethyl bromoacetate, t-butyl bromoaceate, ethyl 3-chloropropionate, ethyl 3-bromopropionate, ethyl 5-bromovalerate, ethyl 4-bromobutyrate, 3-chloropropionamide, 2-bromoethylbenzene, substituted 2-bromoethylbenzene, 1-chloro-3-phenylpropane, 1-bromo-4-phenylbutane, and the like, or nitrogen mustards, including 2-dimethylaminoethyl chloride, 2-diethylaminoethyl chloride, and 3-dimethylaminopropyl chloride. The reaction is carried out in a suitable solvent, such as diethyl ether, tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, or acetonitrile and using a suitable base, such as sodium hydride, potassium hydride, potassium t-butoxide, and lithium diisopropylamide. The reaction is generally carried out at temperatures of xe2x88x9270xc2x0 C. and room temperature and require from about 1-24 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
Alternately, as appreciated by those skilled in the art, an ether formation can also be carried out by a procedure similar to the one above using a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x88x921xe2x80x94CH2OH in which the hydroxy group is first converted to a leaving group, such as chloro, bromo, or mesylate and a suitable alcohol which transfers Q or protected Q as desired in the final product of formula (1), such as benzyl alcohol, substituted benzyl alcohol, phenol, substituted phenol, and the like. The conversion of hydroxy to leaving groups, such as chloro, bromo, and mesylate are well known and appreciated in the art.
For example, in an amine alkylation a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 is contacted with 1 to 10 molar equivalents of a suitable akylating agent to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is xe2x80x94NR8xe2x80x94. The reaction may be carried out after protection of the amine function of R3xe2x80x3 in which R8 is hydrogen by a suitable protecting group, such as benzyl or t-Boc. For an amine alkylation a suitable alkylating agent is one as described above for the ether formation and also includes alkylhalides, such as methyl iodide, methyl bromide, ethyl bromide, propyl bromide, propyl chloride, butyl bromide, butyl chloride, and the like. The reaction is carried out in a suitable solvent, such as methanol, ethanol, dimethylformamide, or pyridine and using a suitable base, such as sodium carbonate, triethylamine, N,N-diisopropylethylamine or pyridine. The reaction is generally carried out at temperatures of room temperature to the refluxing temperature of the solvent and require from about 1-24 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
Alternately, for example, in an amine alkylation a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NH2 is contacted in a reductive alkylation with a suitable aldehyde to give a compound of formula (10) useful as an intermediate for preparing compounds in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is xe2x80x94NR8xe2x80x94. A suitable aldehyde is one which transfers Q or protected Q as desired in the final product of formula (1), such as benzaldehyde and substituted benzaldehydes. The reaction is carried out in a suitable solvent, such as methanol, ethanol, tetrahydrofuran, or mixtures of methanol or ethanol and tetrahydrofuran. The reaction may be carried out in the presence of a drying agent, such as sodium sulfate or molecular sieves. The reaction is carried out in the presence of from 1.0 to 6.0 molar equivalents of a suitable reducing agent, such as, sodium borohydride or sodium cyanoborohydride with sodium cyanoborohydride being preferred. It may be advantageous to maintain the pH in the range of about 4 to 6. The reaction is generally carried out at temperatures of from 0xc2x0 C. to the refluxing temperature of the solvent. Generally, the reactions require 1 to 72 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
For example, in an amido formation a compound of formula (9) in which R3xe2x80x3 is is xe2x80x94(CH2)mxe2x80x94CO2H is contacted with a suitable amine in an amide formation to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is amido. Such amide formation reactions using carboxy activation or suitable coupling agents are well known in the art and described above. A suitable amine, HNR8Q, gives rise to R8 and Q as desired in the final product of formula (1), such as methylamine, ethylamine, propylamine, butylamine, N-methyl benzylamine, benzyl xcex2-alanine, 4-(3-aminopropyl)morpholine, and the like.
For example, in an amide formation a compound of formula (9) in which R3xe2x80x3 is is xe2x80x94(CH2)mxe2x80x94NHR8 is contacted with a suitable carboxylic acid in an amide formation to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is amide. Such amide formation reactions using carboxy activation or suitable coupling agents are well known in the art and described above. Suitable carboxylic acids, QC(O)xe2x80x94OH, are ones give rise to Q as desired in the final product of formula (1), such as benzoic acid, substituted benzoic acids, phenyl acetic acids, substituted phenylacetic acids, mono-t-butyl malonate, and the like.
For example, in a urea formation a compound of formula (9) in which R3xe2x80x3 is is xe2x80x94(CH2)mxe2x80x94NHR8 is contacted with an appropriate isocyanate, Oxe2x95x90Cxe2x95x90N-Q, to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is urea. An appropriate isocyanate is one which gives rise to Q as desired in the final product, such as phenyl isocyanate, substituted phenyl isocyanate, napthyl isocyanate, ethyl isocyanatoacetate, and the like. The reaction is carried out by adding an equivalent of, or a slight molar excess of, an appropriate isocyanate is added to a solution of a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 in a suitable solvent, such as diethyl ether, benzene, or toluene. The reaction is carried out at temperature of from about 0xc2x0 C. to the refluxing temperature of the solvent and require about 1-24 hours. The product can be isolated and purified by techniques well known in the art, such as filtration, extraction, evaporation, trituration, chromatography, and recrystallization.
For example, in an N-carbamoyl formation a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 is contacted with an appropriate chloroformate to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is N-carbamoyl. An appropriate chloroformate is one which gives rise to Q as desired in the final product of formula (1). Examples of chloroformates include benzyl chloroformate, naphthyl chloroformate, phenyl chloroformate, and substituted phenyl chloroformates, such as 4-chlorophenyl chloroformate, 4-methylphenyl chloroformate, 4-bromophenyl chloroformate, 4-fluorophenyl chloroformate, 4-methoxyphenyl chloroformate and the like. The reaction is carried out by adding an equivalent of, or a slight molar excess of, an appropriate chloro formate to a solution of a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 in a suitable solvent, such as toluene, tetrahydrofuran, dimethylformamide, dichloromethane, pyridine, or chloroform. The reaction is carried out in the presence of an excess of a suitable base, such as triethylamine, sodium carbonate, potassium bicarbonate, pyridine or N,N-diisopropylethylamine. The reaction is carried out at a temperature of from xe2x88x9270xc2x0 C. to the refluxing temperature of the solvent and generally requires from 30 minutes to 24 hours. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, chromatography, and recrystallization.
For example, in an O-carbamoyl formation a compound of formula (9) in which R3xe2x80x3 is
xe2x80x94(CH2)mxe2x88x921xe2x80x94CH2OH is contacted with an appropriate isocyanate, as defined above for urea formation, to give a compound of formula (10) in which R3 is xe2x80x94(CH2)m-Z-Q in which Z is O-carbamoyl. The reaction is carried out in a suitable solvent, such as diethyl ether, tetrahydrofuran, dimethylformamide, or acetonitrile. The reaction may be facilitated by the use of catalytic amount of a suitable base, such as sodium hydride, potassium hydride, or potassium t-butoxide. The reaction is generally carried out at temperatures of from xe2x88x9220xc2x0 C. to room temperature and require from about 1-24 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
For example, in a sulfonamide formation to prepare a compound in which R3 is xe2x80x94(CH2)mxe2x80x94NR8xe2x80x2SO2xe2x80x94Y1, a compound of formula (9) in which R3xe2x80x3 is xe2x80x94(CH2)mxe2x80x94NHR8 is contacted with an appropriate sulfonamide forming reagent. An appropriate sulfonamide forming reagent, such as a sulfonyl chloride, Y1S(O)2Cl, or sulfonyl anhydride, Y1(O)2Sxe2x80x94Oxe2x80x94S(O)2 Y1, is one which gives rise to Y1 as desired in the final product. Examples of appropriate sulfonamide forming reagents are, benzenesulfonyl chloride, dansyl chloride, N-morpholinylsulfonyl chloride, N-piperidinylsulfonyl chloride, 2,4,5-trichlorobenzenesulfonyl chloride, 2,5-dichlorobenzenesulfonyl chloride, 2,4,6-triisopropylbenzenesulfonyl chloride, 2-mesitylenesulfonyl chloride, 4-bromobenzenesulfonyl chloride, 4-fluorobenzenesulfonyl chloride, 4-chlorobenzenesulfonyl chloride, 4-methoxybenzenesulfonyl chloride, 4-t-butylbenzenesulfonyl chloride, p-toluenesulfonyl chloride, 2,3,4-trichlorobenzenesulfonyl chloride, 2,5-dimethoxybenzenesulfonyl chloride, 4-ethylbenzenesulfonyl chloride, 3,4-dimethoxybenzenesulfonyl chloride, 2,6-dichlorobenzenesulfonyl chloride, 3-bromobenzenesulfonyl chloride, 4-n-butylbenzenesulfonyl chloride, benzenesulfonic anhydride, 4-toluenesulfonic anhydride, and 2-mesitylenesulfonic anhydride. The reaction is carried out in a suitable solvent, such as tetrahydrofuran, dichloromethane, pyridine, or chloroform and in the presence of an excess of a suitable base, such as triethylamine, sodium carbonate, pyridine, or N,N-diisopropylethylamine. The reaction is carried out at a temperature of from xe2x88x9250xc2x0 C. to the refluxing temperature of the solvent The reaction generally requires from 30 minutes to 24 hours. The product can be isolated and purified by techniques well known in the art, such as extraction, evaporation, chromatography, and recrystalation.
In Reaction Scheme B, optional step 3, a compound of formula (10) in which R3 is as desired in the final product of formula (1) undergoes a selective thiol deprotection to give a compound of formula (11). Such selective thiol deprotections using suitable protecting groups are well known and appreciated in the art as discussed in Reaction Scheme A, step 4, above.
In Reaction Scheme B, step 4, a compound of formula (11) undergoes a modification reaction as described in Reaction Scheme A, step 5, above, to give a compound of formula (7).
In Reaction Scheme B, optional step 5, a compound of formula (10), (11), or (12) is deprotected to give a compound of formula (1) as discussed in Reaction Scheme A, above.
Alternate routes for preparing the compounds of formula (2) in which Y is bromo are presented in Reaction Schemes C.1 and C.2. 
In Reaction Scheme C.1, an appropriate xcex1-amino carboxylic acid of formula (20) is deaminobrominated to give a compound of formula (2) in which Y is bromo. An appropriate xcex1-amino carboxylic acid of formula (20), and protected forms thereof, is one which is one in which R3xe2x80x2 is as described above in Reaction Scheme A, step 8, above. In addition, xcex1-amino carboxylic acid of formula (20) may also be one in which the stereochemistry at the R3xe2x80x2 bearing carbon gives rise after displacement to the stereochemistry as desired at that carbon in the final product of formula (1). Such appropriate xcex1-amino carboxylic acid of formula (20), are commercially available or may be readily prepared by techniques and procedures well known and appreciated by one of ordinary skill in the art. For example, L-alanine, D-alanine, L-valine, D-valine, D-norvaline, L-leucine, D-leucine, D-isoleucine, D-tert-leucine, glycine, L-glutamic acid, D-glutamic acid, L-glutamine, D-glutamine, L-lysine, D-lysine, L-ornithine, D-ornithine, (D)-(xe2x88x92)-2-aminobutyric acid, D-threonine, D-homoserine, D-allothreonine, D-serine, D-2-aminoadipic acid, D-aspartic acid, D-glutamic acid, D-lysine hydrate, 2,3-diaminopropionic acid monohydrobromide, D-ornithine hydrochloride, D,L-2,4-diaminobutyric acid dihydrochloride, L-meta-tyrosine, D-4-hydroxyphenylglycine, D-tyrosine, L-phenylalanine, D-phenylalanine, D,L-2-fluorophenylalanine, beta-methyl-D,L-phenylalanine hydrochloride, D,L-3-fluorophenylalanine, 4-bromo-D,L-phenylaline, L-phenylalanine, L-phenylglycine, D-phenylglycine, D,L-4-fluorophenylalanine, 4-iodo-D-phenylalanine, D-homophenylalanine, D,L-2-fluorophenylglycine, D,L-4-chlorophenylalanine, and the like, are all commercially available and the methods in D. A. Evans, et al. J. Am. Chem. Soc., 112, 4011-4030 (1990); S. Ikegami et al. Tetrahedron, 44, 5333-5342 (1988); W. Oppolzer et al. Tet. Lets. 30, 6009-6010 (1989); Synthesis of Optically Active xcex1-Amino-Acids, R. M. Williams (Pergamon Press, Oxford 1989); M. J. O""Donnell ed.: xcex1-Amino-Acid Synthesis, Tetrahedron Symposia in print, No. 33, Tetrahedron 44, No. 17 (1988); U. Schxc3x6llkopf; PureAppl. Chem. 55, 1799 (1983); U. Hengarter et al. J. Org. Chem., 44, 3748-3752 (1979); M. J. O""Donnell et al. Tet. Lets., 2641-2644 (1978); M. J. O""Donnell et al. Tet. Lets. 23, 4255-4258 (1982); M. J. O""Donnell et al. J. Am. Chem. Soc., 110, 8520-8525 (1988).
The deaminobromination described in Reaction Scheme C.1 can be performed utilizing conditions described in Compagnone, R. S. and Rapoport, H., J. Org. Chem., 51, 1713-1719 (1986); U.S. Pat. No. 5,322,942, issued Jun. 21, 1994; Overberger, C. G. and Cho, I., J. Org. Chem., 33, 3321-3322 (1968); or Pfister, K. et al., J. Am. Chem. Soc., 71, 1096-1100 (1949).
For example, an xcex1-amino carboxylic acid of formula (20) and a suitable bromide, such as hydrogen bromide or potassium bromide in acidic solution, such as sulfric acid, is treated with sodium nitrite. The reaction temperature is carried out a temperatures of from about xe2x88x9225xc2x0 C. to about ambient temperature and require about 1 to 5 hours. The product can be isolated and purified by techniques well known in the art, such as acidification, extraction, evaporation, chromatography, and recrystallization to give the compound of formula (11)in which Y is bromo. The product can be isolated and purified by techniques well known and appreciated in the art, such as acidification, basification, filtration, extraction, evaporation, trituration, chromatography, and recrystallization. 
In Reaction Scheme C.2, an appropriate carboxylic acid of formula (21) is brominated to give compound of formula (2) in which Y is bromo. An appropriate carboxylic acid of formula (21), and protected forms thereof, is one which is one in which R3xe2x80x3 is as defined in Reaction Scheme A, step 8, above. In addition, carboxylic acid of formula (21) may also be one in which the stereochemistry at the R3xe2x80x2 bearing carbon gives rise after displacement to the stereochemistry as desired at that carbon in the final product of formula (1).
For example, a mixture of a carboxylic acid of formula (21) and dry red phosphorous are treated dropwise with bromine at temperature ranging from about xe2x88x9220xc2x0 to about 10xc2x0 C. The reaction mixture is then warmed to room temperature and then heated to about 80xc2x0 C. for about 2-5 hours. The reaction mixture is then cooled to room temperature, poured into water containing sodium bisulfite, and neutralized using solid sodium carbonate. The aqueous layer is extracted and acidified with a suitable acid, such as concentrated hydrochloric acid. The precipitate is collected by filtration and dried to give the compound of formula (2) in which Y is bromo. The product can be isolated and purified by techniques well known and appreciated in the art, such as acidification, basification, filtration, extraction, evaporation, trituration, chromatography, and recrystallization.
Compounds of formula (20) and (21) in which R3xe2x80x2 is a xe2x80x94(CH2)mxe2x80x94W for use in Reaction Schemes C.1 and C.2 are prepared according to Reaction Scheme D.1 and D.2. 
In Reaction Scheme D.1 an appropriate xcfx89-amino carboxylic acid of formula (22) is converted to an compound of formula (21) in which R3xe2x80x2 is Wxe2x80x94(CH2)mxe2x80x94. An appropriate xcfx89-amino carboxylic acid of formula (2) is one in which m is as desired in the final product of formula (1) and are readily available in the art. For example, the reaction is carried out in a suitable polar solvent, such as water, ethanol, diethyl ether, tetrahydrofuran, or a water/ethanol solvent mixture using a suitable base, such as sodium carbonate and N-carbethoxyphthalimide. The reaction mixture is typically stirred at about ambient temperature for 1-5 hours. The product can be isolated and purified by techniques well known in the art, such as acidification, extraction, evaporation, chromatography, and recrystallization to give the desired compound of formula (21) in which R3xe2x80x2 is Wxe2x80x94(CH2)mxe2x80x94. 
Reaction Scheme D.2, step 1, an appropriate xcex1,xcfx89-diamino acid of formula (23) undergoes, a selective N-xcex1-protection to give an N-xcex1-protected-xcfx89-diamino acid of formula (24). An appropriate xcex1,xcfx89-diamino acid of formula (23) is one in which m is as desired in the final product of formula (1).
For example, a selective N-xcex1-protection of a suitable xcex1,xcfx89-diamino acid, such as L-lysine (formula (23) in which m is 4), is accomplished by masking the xcfx89-amino group by formation of a benzylidene imine. The benzylidene imine is formed by dissolving L-lysine monohydrochloride in lithium hydroxide and cooling the solution to a temperature ranging from about 0xc2x0 to 10xc2x0 C. Freshly distilled benzaldehyde is then added and the solution is shaken. N-xcfx89-benzylidene-L-lysine is recovered by filtration and evaporation. The xcex1-amino group of the N-xcfx89-benzylidene-L-lysine then undergoes protection, such as the introduction of a Cbz or t-Boc group, followed by hydrolytic cleavage of the imine in situ to give N-xcex1-benzyloxy-carbonyl-L-lysine. Accordingly, N-xcfx89-benzylidene-L-lysine is added to a mixture of sodium hydroxide and ethanol, cooled to a temperature of from about xe2x88x925xc2x0 to about
xe2x88x9225xc2x0 C. Then, precooled solutions of benzyloxycarbonyl chloride in a solvent, such as ethanol, is added to the reaction mixture. The temperature is maintained in a range of from about xe2x88x9210xc2x0 to about xe2x88x9225xc2x0 C. during the course of addition, and may allowed to rise afterwards. The reaction mixture is then acidified using a suitable acid, such as precooled hydrochloric acid, and N-xcex1-benzyloxycarbonyl-L-lysine, which corresponds to formula (24) where m is 4, is recovered by filtration evaporate and recrystallization.
In Reaction Scheme D.2, step 2, N-xcex1-benzyloxycarbonyl-L-lysine or other compounds of formula (24) is converted to xcfx89-phthalimido-xcex1-benzyloxycarbonyl-L-lysine or other xcfx89-phthalimido-xcex1-aminoprotected carboxylic acid of formula (25) by the method described in Reaction Scheme D.1, above.
In Reaction Scheme D.2, step 3, the xcfx89-phthalimido-xcex1-aminoprotected carboxylic acid of formula (25) is deprotected to give compound of formula (20) in which R3xe2x80x2 is Wxe2x80x94(CH2)mxe2x80x94.
For example, xcfx89-phthalimido-xcex1-benzyloxycarbonyl-L-lysine is contacted with hydrogen in the presence of a hydrogenation catalyst, such as 10% palladium/carbon. The reactants are typically contacted in a suitable solvent mixture such as ethanol, methanol, water, ethanol/water mixtures, or methanol/water mixtures. The reactants are typically shaken under a hydrogen atmosphere of 35-45 psi at room temperature for a period of time ranging from 5-24 hours. The product is typically recovered by filtration and evaporation of the solvent.
A route for preparing the compounds of formula (2) in which Y is protected thio is presented in Reaction Scheme F. The reagents and starting materials are readily available to one of ordinary skill in the art. In Reaction Scheme H all substituents, unless otherwise indicated, are as previously defined. 
In Reaction Scheme F, step 1, a bromoacetate of formula (26) is contacted with an appropriate thiol to give a protected acetic acid ester of formula (27). In a bromoacetate of formula (26) Pg5 is a protecting group, such as methyl, ethyl, t-butyl, and benzyl. An appropriate thiol is one which gives rise to a protected thio group, Y, in the product of formula (11). The use of 4-methoxybenzylmercaptan is preferred.
For example, a bromoacetate of formula (26) is contacted with an appropriate thiol in a suitable organic solvent, such as dimethylformamide. Advantageously, the solvent is degassed. The reaction is carried out using a suitable base, such as sodium hydroxide, triethylamine, or N,N-diisopropylethylamine. The reaction is carried out at temperatures of from about xe2x88x9250xc2x0 to about ambient temperature and requires about 1 to 72 hours. The protected acetic acid ester of formula (27) can be isolated and purified by methods well known and appreciated in the art, such as extraction, evaporation, chromatography, and distillation, and recrystallization.
In Reaction Scheme F, step 2, the protected acetic acid ester of formula (27) is alkylated with an appropriate akylating agent to give a compound of formula (28). In Reaction Scheme F, step 2, an appropriate alkylating agent is one which transfers R3xe2x80x2 which is R3 as desired in the final product of formula (1) or gives rise after deprotection to R3 as desired in the final product of formula (1) or gives rise to R3xe2x80x3 as defined in Reaction Scheme B, step 1. Appropriate alkylating agents include alkylhalides, such as methyl iodide, methyl bromide, ethyl bromide, propyl bromide, propyl chloride, butyl bromide, butyl chloride, and the like; benzyl bromide, benzyl chloride, substituted benzyl bromide, substituted benzyl chloride, ethyl bromoacetate, t-butyl bromoaceate, ethyl 3-chloropropionate, ethyl 3-bromopropionate, ethyl 5-bromovalerate, ethyl 4-bromobutyrate, 3-chloropropionamide, 2-bromoethylbenzene, substituted 2-bromoethylbenzene, 1-chloro-3-phenylpropane, 1-bromo-4phenylbutane, and the like, N-(2-bromoethyl)phthalimide, N-(3-bromopropyl)phthalimide, N-(4-bromobutyl)phthalimide, and the like; 1-bromo-2-phenylethane, 1-bromo-3-phenylpropane, 1-bromophenylbutane, and the like.
For example, a protected acetic acid ester of formula (27) is alkylated with an appropriate alkylating agent. The reaction is carried out in a suitable solvent, such as diethyl ether, tetrahydrofuran, dimethylformamide, and toluene using a suitable base, such as sodium hydride, potassium hydride, potassium t-butoxide, lithium bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide, potassium bis(trimethylsilyl)amide, or lithium diisopropylamide. The reaction is generally carried out at temperatures of about xe2x88x9270xc2x0 C. to about room temperature and require from about 1-24 hours. The product can be isolated by techniques well known in the art, such as extraction, evaporation, and precipitation and can be purified by chromatography and recrystallization.
In Reaction Scheme F, step 3, the compound of formula (28) the carboxy protecting group Pg5 is selectively removed to give a compound of formula (3b) in which Y is protected thio. Such deprotection of esters to acids in the presence of suitable thio protecting groups are well known and appreciated in the art.
A general preparation of compounds of formula (3) is set forth in Reaction Scheme G. 
In Reaction Scheme G, step 1, an appropriate compound of structure (30) is functionalized to give the corresponding compound of structure (31). An appropriate compound of structure (30) in one in which R2 is as desired in the final product of formula (1).
For example, an appropriate compound of formula (30) is treated initially with lithium chloride and a suitable non-nucleophilic base, such as collidine in a suitable solvent such as dimethylformamide. This is followed by treatment with a suitable mesylating agent, such as mesyl chloride. The reaction is typically carried out at a temperature range of from xe2x88x9230xc2x0 C. to room temperature, preferably 0xc2x0 C. and for a period of time ranging from 2-10 hours. The product is recovered from the reaction mixture by extractive methods as are known in the art and may be purified by chromatography.
In Reaction Scheme G, step 2, the methanesulfonate functionality of the compound of formula (31) is eliminated and the chloro substituted with iodo to a compound of formula (32).
For example, a compound of formula (31) is treated with a suitable non-nucleophilic base, such as potassium tert-butoxide in a suitable aprotic organic solvent, such as diethyl ether. The reaction is typically carried out at a temperature range of from xe2x88x9230xc2x0 C. to room temperature, preferably 0xc2x0 C. and for a period of time ranging from 15 minutes to hours to give the corresponding 1-chloromethyl-2-vinyl-benzene derivative which is recovered from the reaction mixture by extractive methods as are known in the art. The 1-chloromethyl-2-vinyl-benzene derivative is then treated with a suitable iodinating agent, such as sodium iodide, in a suitable solvent, such as acetone. The reaction is carried out at a temperature range of from room temperature to reflux temperature of the solvent and for a period of time ranging from 15 minutes to hours. The product is recovered from the reaction mixture by extractive methods as are known in the art.
In Reaction Scheme G, step 3, a compound of formula (32) is subjected to an addition, elimination reaction with 2-(bis-methylsulfonyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thia-4-aza-tricyclo(5.2.1.0 1,5)dec-4-yl)-ethanone to give the corresponding compound of formula (33).
For example, the anion of 2-(bis-methylsulfonyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thia-4-aza-tricyclo(5.2.1.0 1,5)dec-4-yl)-ethanone is formed by treating 2-(bis-methylsulfonyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thia-4-aza-tricyclo(5.2.1.0 1,5)dec-4-yl)-ethanone with a suitable non-nucleophilic base, such as n-butyllithium in a suitable aprotic organic solvent, such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9278xc2x0 C. to xe2x88x9230xc2x0 C., preferable xe2x88x9278xc2x0 C. and for a period of time ranging from 30 minutes to 2 hours. A compound of formula (32) is then added and the reaction is carried out at a temperature range of from xe2x88x9278xc2x0 C. to room temperature for a period of time ranging from 1-24 hours. The product is recovered from the reaction mixture by extractive methods as are known in the art and may be purified by chromatography.
In Reaction Scheme G, step 4, a compound of formula (33) is hydrolyzed to give a compound of formula (34).
For example, a compound of formula (33) is treated with a suitable acid such as aqueous hydrochloric acid in a suitable organic solvent such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to room temperature and for a period of time ranging from 30 minutes to 20 hours. Evaporation of the solvent followed by treatment with inorganic base such as aqueous lithium hydroxide in a suitable organic solvent, such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to room temperature and for a period of time ranging from 30 minutes to 10 hours. After acidification, the corresponding 2-amino-3-(2-vinyl-phenyl)-propionic acid derivative of formula (34) was isolated by evaporation of solvents.
In Reaction Scheme G, step 5, the amino functionality of a compound of formula (34) is protected to give the corresponding compound of formula (35).
For example, a compound of formula (34) is treated with an appropriate phthalimide protecting agent, such as N-carbethoxyphthalimide in the presence of a suitable non-nucleophilic base, such as aqueous sodium carbonate. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to room temperature and for a period of time ranging from 1-10 hours. The product is recovered from the reaction zone by extractive methods as are known in the art and may be purified by chromatography.
In Reaction Scheme G, step 6, the carboxylic acid functionality of a compound of formula (35) is esterified to give a compound of formula (36).
For example, a compound of formula (35) is treated with 2-(trimethylsilyl)ethanol in the presence of a suitable non-nucleophilic base, such as pyridine, in a suitable organic solvent, such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9230xc2x0 C. to room temperature and for a period of time ranging from 30 minutes to 2 hours. A coupling agent, such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) is then added and the reaction carried out at a temperature range of from xe2x88x9230xc2x0 C. to room temperature for a period of time ranging from 10-48 hours. The product is recovered from the reaction zone by extractive methods as are known in the art and may be purified by chromatography.
In Reaction Scheme G, step 7, the vinyl functionality of a compound of formula (36) is oxidized to give a compound of formula (37).
For example, a compound of formula (36) is treated with ozone in a suitable organic solvent such as methylene chloride and methanol. The reaction is carried out at a temperature range of from xe2x88x9278xc2x0 C. to xe2x88x9250xc2x0 C. and for a period of time necessary for a blue color to persist. After purging the reaction with nitrogen and quenching by methods known in the art, such as addition of dimethylsulfide and pyridine, the product is recovered from the reaction zone by extractive methods as are known in the art and may be purified by chromatography.
In Reaction Scheme G, step 8, a compound of formula (37) is subjected to reductive amination with an appropriate amino acid, tert-butyl ester derivative to give a compound of formula (39). In Reaction Scheme G, step 8, an appropriate amino acid, tert-butyl ester derivative is one in which R1 is as desired in the final product of formula (1) or gives rise to R1 as is desired in the final product of formula (1). In addition, an appropriate amino acid, tert-butyl ester derivative is one in which the stereochemistry is as desired in the final product of formula (1). As is appreciated by those skilled in the art, the use of amino acid, tert-butyl ester derivatives give rise to compounds of formula (3) in which Axe2x80x2 is xe2x80x94O-tert-butyl. It is also understood that appropriate amino acid derivatives can have a variety of carboxy substituents give rise to compounds of formula (3) in which Axe2x80x2 is other than xe2x80x94O-tert-butyl.
For example, a compound of formula (37) is treated with an appropriate amino acid, tert-butyl ester derivative in an appropriate polar organic solvent, such as methanol under dehydrating conditions, such as molecular sieves. The reaction is carried out at a temperature ranged of from xe2x88x9210xc2x0 C. to reflux temperature of the solvent, preferably room temperature, and for a period of time ranging from 30 minutes to 10 hours. A suitable reducing agent, such as sodium cyanoborohydride, is then added and the reaction is carried out at a temperature range of from xe2x88x9210xc2x0 to reflux temperature of the solvent, preferably room temperature, and for a period of time ranging from 30 minutes to 24 hours. The product is recovered from the reaction zone by extractive methods as are known in art and may be purified by chromatography. As one skill in the art would realize, those amino acid, tert-butyl ester derivatives wherein R1 has a reactive functionality, the reactive functionality may be protected prior to the reductive amination reaction of step 8. The selection and utilization of suitable protecting groups is well known by one of ordinary skill in the art and is described in Protective Groups in Organic Synthesis, Theodora W. Greene, (Wiley 1981).
In Reaction Scheme G, step 9, the ester functionality of a compound of formula (39) is hydrolyzed to give a compound of formula (40).
For example, a compound of formula (39) is treated with an appropriate fluoride reagent, such as tetrabutylammonium fluoride in a suitable organic solvent, such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to room temperature and for a period of time ranging from 30 minutes to hours. The product is recovered from the reaction zone by extractive methods as are known in art and may be purified by chromatography.
In Reaction Scheme G, step 10, a compound of formula (39) is subjected to a ring closure amination reaction to give a compound of formula (40).
For example, a compound of formula (40) is treated with a suitable activating agent, such as isobutylchloroformate, in the presence of a suitable non-nucleophilic base, such as N-methylmorpholine in a suitable organic solvent, such as tetrahydrofuran. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to reflux temperature of the solvent and for a period of time ranging from 30 minutes to 10 hours. The product is recovered from the reaction zone by evaporation and may be purified by chromatography.
In Reaction Scheme G, step 11, the protecting group of a compound of formula (40) is removed to give a compound of formula (3) in which Axe2x80x2 is xe2x80x94O-tert-butyl as the 4-position (S)-isomer.
For example, the phthalimide protecting groups of a compound of formula (40) can be removed using hydrazine monohydrate in a suitable protic solvent such as methanol. The reaction is carried out at a temperature range of from xe2x88x9210xc2x0 C. to room temperature and for a period of time ranging from 2 hours to 4 days. The product is recovered from the reaction zone by filtration and evaporation.
The following examples present typical syntheses as described in the Reaction Schemes above. These examples and preparations are understood to be illustrative only and are not intended to limit the scope of the invention in any way.
Dissolve homophthalic acid (22.2 g, 0.123 mmol) in tetrahydrofuran (250 mL) and add dropwise at room temperature to a slurry of lithium aluminum hydride (15.5 g, 0.407 mol) in tetrahydrofuran (500 mL). Heat at reflux for 18 hours, cool in an ice bath and carefully add, by dropwise addition, water (16 mL), followed by 50% sodium hydroxide (16 mL). Remove the ice bath, add water slowly with stirring and stir until the gray precipitate turns white and evolution of gas ceases. Filter, wash solids with methylene chloride, dry (MgSO4) and evaporate the solvent in vacuo to give 2-(2-hydroxymethyl-phenyl)-ethanol as a viscous oil (18.4 g, 98%).
Mix 2-(2-hydroxymethyl-phenyl)-ethanol (12.0 g, 78.8 mmol) and collidine (23 mL, 0.17 mol) and treat with lithium chloride (7.35 g, 0.173 mmol) in dimethylformamide (125 mL). Cool in an ice bath and treat, by dropwise addition, with mesyl chloride (13.4 mL). Stir at 0xc2x0 C. for 4 hours, partition between ice water (300 mL) and a 1:1 mixture of ether:pentane (2xc3x97400 mL). Wash the organic layer with a saturated solution of CuSO4 (2xc3x97200 mL), dry (MgSO4) and purify by silica gel chromatography (2.5:1 hexane/ethyl acetate followed by 2:1 hexane/ethyl acetate followed by 3:2 hexane/ethyl acetate) to give methanesulfonic acid 2-(2-chloromethyl-phenyl)-ethyl ester as a pale yellow oil (8.8 g, 45%).
Dissolve methanesulfonic acid 2-(2-chloromethyl-phenyl)-ethyl ester (8.8 g, 35.4 mmol) in ether (80 mL) and cool to xe2x88x9235xc2x0 C. Add potassium t-butoxide (10 g, 89 mmol) and stir for 30 minutes. Add water (50 mL) and ether (150 mL), extract, dry (Na2SO4) and purify by silica gel chromatography (2:3 methylene chloride/pentane) to give 1-chloromethyl-2-vinyl-benzene as a colorless oil (4.43 g, 82%).
Dissolve 1-chloromethyl-2-vinyl-benzene (4.0 g, 26 mmol) in acetone (100 mL) and add sodium iodide (4.5 g, 30 mmol). Heat at gentle reflux for 30 minutes. Cool, add water (150 mL) and extract with pentane (200 mL). Dry (MgSO4) and evaporate the solvent in vacuo to give 1-iodomethyl-2-vinyl-benzene (95%).
Dissolve 2-(bis-methylsulfanyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thia-4-aza-tricyclo(5.2.1.0 1,5)dec-4-yl)-ethanone (7.91 g, 21.0 mmol) in tetrahydrofuran (100 mL) and cool to xe2x88x9278xc2x0 C. Treat, by dropwise addition, with 1.6 M n-butyllithium in hexane (13.1 mL, 21 mmol). Stir for 1.5 hours, then add hexamethylphosphotriamide (HMPA) (4.25 mL, 24.4 mmol). Stir for 15 minutes and add, via cannula, a solution of 1-iodomethyl-2-vinyl-benzene (6.1 g, 25 mmol) in tetrahydrofuran (100 mL). Stir overnight at room temperature, partition between saturated ammonium chloride (2xc3x9775 mL) and ethyl acetate (400 mL). Dry (Na2SO4) and purify by silica gel chromatography (2.5:1 hexane/ethyl acetate) to give 2-(bis-methylsulfanyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thia-4-aza-tricyclo(5.2.1.0 1,5)dec-4-yl)-3-(2-vinyl-phenyl)-propan-1-one as a white solid (4.5 g).
Dissolve 2-(bis-methylsulfanyl-methyleneamino)-1-(10,10-dimethyl-3,3-dioxo-3-thiaffaza-tricyclo(5.2.1.0 1,5)dec-4-yl)-3-(2-vinyl-phenyl)-propan-1-one (5.21 g, 10.6 mmol) in tetrahydrofuran (100 mL) and) and 0.75 M hydrochloric acid (100 mL). Stir at room temperature for 24 hours, evaporate the solvent in vacuo to give the hydrochloride salt as a white solid. Dissolve in tetrahydrofuran (200 mL) and water (50 m), add lithium hydroxide monohydrate (1.9 g, 4.5 mmol) and stir at room temperature under a nitrogen atmosphere for 4 hours. Extract into methylene chloride (200 mL) and wash with 2N sodium hydroxide (50 mL). Acidify to pH 2-3 while cooling in an ice bath and concentrate in vacuo to give 2-Amino-3-(2-vinyl-phenyl)-propionic acid as an off-white solid (3.40 g, 100%).
Dissolve 2-amino-3-(2-vinyl-phenyl)-propionic acid (3.40 g) in water (75 mL) and add sodium carbonate (1.97 g, 18.6 mmol) and N-carbethoxyphthalimide (2.81 g, 12.8 mmol). Stir for 2.5 hours, wash with methylene chloride (200 mL), acidify to pH 1 with cold concentrated hydrochloric acid and extract with ethyl acetate (3xc3x97200 mL), dry (Na2SO4), evaporate the solvent in vacuo and purify by silica gel chromatography (1:1:0.02 hexane/ethyl acetate/acetic acid) followed by recrystallization (isopropanol) to give 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-vinyl-phenyl)-propionic acid as a pale yellow solid (2.47 g).
Dissolve 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-vinyl-phenyl)-propionic acid (2.47 g, 7.69 mmol) in tetrahydrofuran (35 mL) and cool in an ice bath. Treat with pyridine (1.6 mL, 20 mmol) and 2-(trimethylsilyl)ethanol (2.3 mL, 16 mmol). Stir for 30 minutes and add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (2.21 g, 11.5 mmol). Stir for 22 hours at 5xc2x0 C., then at room temperature for 1.5 hours. Cool to 0xc2x0 C., add 0.6 times all reagents and stir at room temperature overnight. Dilute with ethyl acetate (150 mL), wash with 5% sulfuric acid (40 mL) and saturated sodium hydrogen carbonate (40 mL). Back extract with methylene chloride (100 mL), wash with brine (30 mL) and dry (Na2SO4). Evaporate the solvent in vacuo and purify by silica gel chromatography (2:1 hexane/ethyl acetate) to give 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-vinyl-phenyl)-propionic acid, 2-trimethylsilanyl-ethyl ester (2.61 g, 81%).
Dissolve 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-vinyl-phenyl)-propionic acid, 2-trimethylsilanyl-ethyl ester (2.61 g, 6.19 mmol) in methylene chloride (70 mL) and methanol (75 mL). Cool to xe2x88x9278xc2x0 C. and treat with ozone until a blue color persists. Purge with nitrogen and add dimethylsulfide (7 mL) and pyridine (0.35 mL). Allow to warm to room temperature gradually overnight. Partition between methylene chloride (100 mL) and water (40 mL). Extract the aqueous with methylene chloride (50 mL), dry (Na2SO4) and purify by silica gel chromatography (2.5:1 hexane/ethyl acetate) to give the title compound as a colorless viscous oil (2.65 g, 100%). Step h: 2-(2-(2-(1,3,-Dioxo-1,3,dihydro-isoindol-2-yl)-2-(2-trimethylsilanyl-ethoxycarbonyl)-ethyl)-benzylamino)-4-methyl-valeric acid, tert-butyl ester.
Dissolve 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-formyl-phenyl)-propionic acid, 2-trimethylsilanyl-ethyl ester (250 mg, 0.590 mmol) in methanol (15 mL) and treat with L-leucine tert-butyl ester hydrochloride (0.66 g, 3.0 mmol). Stir at room temperature for 2 hours with 3A molecular sieves, add sodium cyanoborohydride (0.6 mL of a 1.0M solution in tetrahydrofuran, 0.6 mmol), stir for 0.5 hours, add additional sodium cyanoborohydride (0.3 mL) and stir for 5 hours. Filter through filter aid, evaporate the solvent in vacuo and partition the residue between methylene chloride (100 mL) and saturated sodium hydrogen carbonate (40 mL). Dry (Na2SO4), evaporate the solvent in vacuo and purify by silica gel chromatography (5:1 hexane/ethyl acetate followed by 3:1 hexane/ethyl acetate) to give 2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-(2-formyl-phenyl)-propionic acid, 2-trimethylsilanyl-ethyl ester (221 mg, 63%).
Dissolve 2-(2-(2-(1,3-dioxo-1,3,dihydro-isoindol-2-yl)-2-(2-trimethylsilanyl-ethoxycarbonyl)-ethyl)-benzylamino)-4-methyl-valeric acid, tert-butyl ester (221 mg, 0.372 mmol) in tetrahydrofuran (5 mL) and treat with tetrabutylammonium fluoride (0.43 mL of a 1.0M solution in tetrahydrofuran, 0.43 mmol). Stir for 1.5 hours, evaporate the solvent in vacuo and dissolve the residue in ethyl acetate (75 mL). Wash with 1N hydrochloric acid (25 mL) and brine (25 mL). Dry (Na2SO4) and evaporate the solvent in vacuo to 2-(2-(2-carboxy-2-(1,3-dioxo-1,3,dihydro-isoindol-2-yl)-ethyl)-benzylamino)-4-methyl-valeric acid, tert-butyl ester as a white solid (188 mg).
Dissolve 2-(2-(2-carboxy-2-(1,3-dioxo-1,3,dihydro-isoindol-2-yl)-ethyl)-benzylamino)-4-methyl-valeric acid, tert-butyl ester (188 mg) in tetrahydrofuran (10 mL) and cool in an ice bath. Add sequentially, N-methylmorpholine (86 xcexcL, 0.78 mmol), and isobutylchloroformate (55 xcexcL, 0.43 mmol). Stir for 2 hours, filter, wash salts with dry tetrahydrofuran, evaporate the solvent in vacuo and purify by silica gel chromatography (1:1 hexane/ethyl acetate) to give 2-(4-((1,3-dioxo-1,3,dihydro-isoindol-2-yl)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, tert-butyl ester as a white solid (64 mg, 93%).
Dissolve 2-(4((1,3-dioxo-1,3,dihydro-isoindol-2-yl)-3-oxo-1,3,4,5-tetrahydro-benzo[c]azepin-2-yl)-4-methyl-valeric acid, tert-butyl ester (160 mg, 0.336 mmol) in methanol (3 mL) and treat with a solution of hydrazine monohydrate (0.40 mL, 0.40 mmol) in methanol. Stir at room temperature for 65 hours, filter through filter aid, wash with methylene chloride, filter through filter aid and dry (MgSO4). Evaporate the solvent in vacuo to give the title compound (93 mg, 80.2% for this step).